title: Molecular and genetic analyses of Tic20 homologues in Arabidopsis thaliana chloroplasts.
abstract: The Tic20 protein was identified in pea (Pisum sativum) as a component of the chloroplast protein import apparatus. In Arabidopsis, there are four Tic20 homologues, termed atTic20-I, atTic20-IV, atTic20-II and atTic20-V, all with predicted topological similarity to the pea protein (psTic20). Analysis of Tic20 sequences from many species indicated that they are phylogenetically unrelated to mitochondrial Tim17-22-23 proteins, and that they form two evolutionarily conserved subgroups [characterized by psTic20/atTic20-I/IV (Group 1) and atTic20-II/V (Group 2)]. Like psTic20, all four Arabidopsis proteins have a predicted transit peptide consistent with targeting to the inner envelope. Envelope localization of each one was confirmed by analysis of YFP fusions. RT-PCR and microarray data revealed that the four genes are expressed throughout development. To assess the functional significance of the genes, T-DNA mutants were identified. Homozygous tic20-I plants had an albino phenotype that correlated with abnormal chloroplast development and reduced levels of chloroplast proteins. However, knockouts for the other three genes were indistinguishable from the wild type. To test for redundancy, double and triple mutants were studied; apart from those involving tic20-I, none was distinguishable from the wild type. The tic20-I tic20-II and tic20-I tic20-V double mutants were albino, like the corresponding tic20-I parent. In contrast, tic20-I tic20-IV double homozygotes could not be identified, due to gametophytic and embryonic lethality. Redundancy between atTic20-I and atTic20-IV was confirmed by complementation analysis. Thus, atTic20-I and atTic20-IV are the major functional Tic20 isoforms in Arabidopsis, with partially overlapping roles. While the Group 2 proteins have been conserved over approximately 1.2 billion (1.2 × 10(9) ) years, they are not essential for normal development.
authors.affiliation: Department of Biology, University of Leicester, University Road, Leicester LE17RH, UK.
authors.forename: Ali Reza
authors.initials: AR
authors.lastname: Kasmati
authors.forename: Mats
authors.initials: M
authors.lastname: Töpel
authors.forename: Ramesh
authors.initials: R
authors.lastname: Patel
authors.forename: Ghulam
authors.initials: G
authors.lastname: Murtaza
authors.forename: Paul
authors.initials: P
authors.lastname: Jarvis
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D002736:Chloroplasts / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D016384:Consensus Sequence; D019143:Evolution, Molecular; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D005816:Genetic Complementation Test; D006720:Homozygote; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D016254:Mutagenesis, Insertional; D009154:Mutation; D056771:Ovule / Q000254:growth & development; D010802:Phylogeny; D012639:Seeds / Q000254:growth & development; D014162:Transfection; D019076:Transgenes
chemical_list: D029681:Arabidopsis Proteins; D026901:Membrane Transport Proteins; C457764:Tic20 protein, Arabidopsis